Channels: Where Disciplines Meet Fall 2021 Volume 6, Number 1
Editorial Board Julie Deardorff Director of Library Collection Services; Associate Professor of Library Science, Cedarville University Greg Martin Digital Commons Director; Associate Professor of Library Science, Cedarville University Dr. Mark Owens Assistant Professor of New Testament Theology, Cedarville University Jennifer Wingerter Assistant Professor of Professional Writing and Information Design, Cedarville University Sophia Cali Junior Professional Writing and Information Design Student, Cedarville University Elisabeth Carlsen Sophomore Professional Writing and Information Design Student, Cedarville University Brianna DeMan Sophomore Professional Writing and Information Design Student, Cedarville University Channels, a bi-annual undergraduate research journal, consists of faculty sponsored articles from various disciplines across Cedarville University’s campus. Each article is sponsored by a faculty member and will be published in digital and print format. Channels strives to provoke thoughtful discussion among readers. Authors explore new ideas, generate creative solutions to existing problems, and develop knowledge in new ways. DigitalCommons@Cedarville provides a publication platform for fully open access journals, which means that all articles are available on the Internet to all users immediately upon publication. However, the opinions and sentiments expressed by the authors of articles published in our journals do not necessarily indicate the endorsement or reflect the views of DigitalCommons@Cedarville, the Centennial Library, or Cedarville University and its employees. The authors are solely responsible for the content of their work. Please address questions to dc@cedarville.edu. ISSN: 2474-2651
Table of Contents Fall 2021 Volume 6, Number 1 Differential MicroRNA Expression in Glioblastoma as a Therapeutic Target 1 or Potential Biomarker Drew Hines, Levi Marcum, Aubrey Strong, Ryan Wade Humor in the Foreign Language Classroom 9 Emily Walker
Channels • 2021 • Volume 6 • Number 1 Page 1 Channels Vol. 6 No. 1 (2021): 1–8 ISSN 2474-2651 © 2021, Hines, Marcum, Strong, Wade, licensed under CC BY-NC-ND (http://creativecommons.org/licenses/by-nc-nd/4.0/) Differential MicroRNA Expression in Glioblastoma as a Therapeutic Target or Potential Biomarker Drew Hines, Levi Marcum, Aubrey Strong, Ryan Wade Science and Mathematics Introduction Function of miRNA type of short non-coding RNA of about 20–25 nucleotides, microRNA (miRNA), are found in most eukaryotic cells and certain viruses. MiRNA is primarily involved in the posttranscriptional regulation of gene expression, and they mainly operate by inhibiting mRNA translation or cleaving/destabilizing. RNA polymerase II transcribes Pre-miRNA, which is then cleaved in succession by the proteins Drosha and Dicer. Once the miRNA is fully mature, it can do two things. First, it can bind with the RNA-induced silencing complex, which orients the miRNA so it can more easily interact with mRNA. When the complex recognizes a specific sequence of nucleotides on the mRNA, it can either directly or indirectly cleave the strand. Second, the miRNA can negatively regulate mRNA through a negative feedback loop or a sort of feed-forward loop. Due to the abundance of miRNA and the existence of extracellular circulating miRNA—miRNA that is released into body fluids like blood and cerebrospinal fluid—miRNA has the potential to be an effective biomarker for many diseases. Glioblastoma Glioblastoma is one of the most lethal human cancers. With less than 5% of patients surviving past 5 years, glioblastoma ranks as the most common of primary malignant central nervous system tumors. Doctors can currently treat the tumor through gross total surgery resection followed by radiotherapy and temozolomide, and tumor-treating fields is a new method proven to extend survival (Gimple et al., 2019). Poor prognosis is typical for many glioblastoma patients due to the complexity of the tumor. High degrees of intramural cellular heterogeneity, the infiltrative and migratory nature of glioblastoma cells, and a high rate of tumor recurrence all contribute to its poor prognosis. Recurrent tumors also display distinct divergences from the original tumor, which severely inhibits the information obtained from initial biopsies (Gimple et al., 2019). Another contribution to poor prognosis is the presence of differentially expressed microRNAs. Malignancy and stemness-associated miRNAs have been identified in glioblastoma and may regulate genes associated with cancer development and
Page 2 Hines, Marcum, Strong, Wade • Differential MicroRNA Expression radio resistance (Gimple et al., 2019). As a key component in glioblastoma’s resistance to modern therapies, miRNAs are ideal targets for the progression of glioblastoma treatment effectiveness. MicroRNA in Other Cancers Besides being found in glioblastoma, microRNA (miRNA) also has a distinct role in other cancers. MicroRNA is the key to knowing how cancer stem cells (CSCs), specifically neural CSCs, are created and can ultimately be destroyed because of how CSCs downregulate pro-apoptotic miRNA as compared to normal neural stem cells (NSCs) (Diana et al., 2020). This allows for unregulated growth of the cancerous cells and leads to a multitude of possible cancers. When restored, these pro-apoptotic miRNAs could inhibit anti-apoptotic genes allowing them to act as tumor suppressant miRNAs (Diana et al., 2020). This kind of information is important to treat multiple different neural cancers. CSCs avoid apoptosis and are nearly immune to most regular forms of treatment (Diana et al., 2020). Neural CSCs can escape apoptosis by the downregulation of miRNAs and avoid other treatments by downregulation of death receptors or anti-apoptotic factors. CSCs including brain tumor stem cells, glioma stem cells, medulloblastoma, neuroblastoma, and melanoma stem cells are the cause of multiple different cancers. (Diana et al., 2020). Although other cancers use miRNAs, the focus in research has been on the neurological side. MicroRNA and Autoimmune Diseases MicroRNA plays an important role in the development or regulation of cancer, but recent studies have also shown correlations between the dysregulation of microRNA and the development of autoimmune diseases. Autoimmune diseases are a class of disorders that occur when the body develops an immune response to self-antigens. T cells, part of the immune system, change with the environment and can have a significant impact on autoimmunity. T helper type 17 (Th17) cells release cytokines that activate parts of the immune system such as macrophages and neutrophils (Liu et al., 2018). This increases inflammation and immune response throughout the body. However, there are two types of regulatory T (Treg) cells that have the opposite effect. Naturally occurring Treg (nTreg) cells inhibit inflammation and autoimmunity through cell communication, and inducible Treg (iTreg) cells activate cytokines that also have suppressive properties (Liu et al., 2018). In order to maintain homeostasis in the human body, it is important that Th17 cells and Treg cells are balanced. Upregulation of the gene that codes for Th17 causes increased inflammation and an increased risk of developing autoimmune diseases. Similarly, the downregulation of genes that code for Treg cells also causes increased autoimmunity. The relationship between these two types of T cells is crucial for the human body to function properly. MicroRNA also plays a role in the gene regulation of these two cell types, therefore impacting their balance and overall autoimmunity. Upregulation of certain microRNAs inhibits tensin homolog and phosphatases, the result highly activated T cells in immune response. MiR-214 and miR-182 are specifically known for having this effect (Colamatteo et al., 2019). MiR-155 seems to play a role in the suppression and activation of cytokines, and studies in mice have shown evidence that downregulation of miR-146 causes both inflammation autoimmunity to increase, as well as an overall suppression of T cells (Colamatteo et al., 2019). Altogether, an upregulation of miRNAs that control the response of Th17 can lead to chronic inflammation and autoimmunity. Similarly,
Channels • 2021 • Volume 6 • Number 1 Page 3 downregulation of miRNAs that control the Treg response also causes inflammation and autoimmunity to increase. Heightened inflammation and dysregulated immune response lead to development of autoimmune disorders such as rheumatoid arthritis, multiple sclerosis, psoriasis, inflammatory bowel disease, and systemic lupus (Liu et al., 2018). Analysis MicroRNA Expression in Healthy Astrocytes MicroRNA (miRNA) plays an important part in maintaining the normal functions of the central nervous system (Sun et al., 2019), and they often have an inhibitory function. Although its role is still not completely clear, an accumulating collection of evidence indicates that miRNA is a key part of many of the roles and tasks of the astrocyte. For instance, miRNA seems to be necessary in the formation of synapses. Research has shown that increased levels of CCL5, a target of miRNA, caused deficiencies in synapse formation (Sun et al., 2019). In addition, this research further supported the theory that miRNA plays a pivotal role in regulating the activation of astrocytes. When an astrocyte becomes activated, it gains the ability to secrete various signaling substances that regulate neuron development, function, and connectivity (Jovicic & Gitler, 2017). Among these substances are exosomes carrying miRNA shown to be significantly different from the miRNA that remains inside the cell. The miRNA carried by exosomes provide a mechanism for differentiating the function of various miRNAs within the cell. Also, while not much is yet known about this exosomal miRNA, recent evidence shows that miRNA plays a key role in the inflammatory response, diminishing the activity of target neurons and downregulating the transcription of proteins required for neuronal excitability (Chaudhuri et al., 2018). In fact, miRNA likely plays a major role in the neurogenic stress response (Luarte et al., 2017). For this reason, certain miRNA could be a good biomarker for central nervous system inflammation and stress points (Lafourcade et al., 2016). Finally, there is no one fast and easy way to tell what a healthy expression of miRNA in astrocytes is since the answer will depend on many factors, such as age and anatomical location. Researchers find a high expression of miRNA in the fetal germinal matrix, likely for developmental purposes (Rao et al., 2016). There is a higher expression of miRNA in adult white matter than in fetal white matter, while in grey matter the two seem comparable. Also, similar overall levels of miRNA exist between adult white and grey matter; however, different levels of various miRNA appear to be expressed in each (Rao et al., 2016). MicroRNA Expression in Glioblastoma MiRNAs have a close relation to the biological features of the glioblastoma stem cells (GSCs) and predict the survival in glioblastoma patients. Fifty-one miRNAs are associated with glioblastoma’s stem-like phenotype, and nine were identified to be strongly upregulated in GSCs: miR-9-3p, miR93-3p, miR-93-5p, miR-106b-5p, miR-124-3p, miR-153-3p, miR-301a-3p, miR-345-5p, and miR652-3p (Sana et al., 2018). Both miR-9-3p and the hairpin counterpart miR-9-5p are expressed in the brain. These miRNAs affect the Notch signaling pathways, which surprisingly promotes differentiation of neural stem cells. Furthermore, this affected Notch pathway appears to induce
Page 4 Hines, Marcum, Strong, Wade • Differential MicroRNA Expression miR-9/9*. (An asterisk is added to the name of the miRNA strand to denote the star strand.) Abundant in CD133+ GSCs, these miRNAs seem to contribute to glioblastoma’s high resistance to modern therapy (Sana et al., 2018). The cluster containing miR-106-5p, miR-93-5p, and miR-93-3p seems to have a close connection to the biology of stem cells. Research shows the inhibition of this cluster in CD44+ gastric cancer stem-like cells suppresses the TGF-ß/Smad pathway (Sana et al., 2018). MiR-153 was shown to have a high expression in GSCs, and its overexpression reduced GSC’s tumorigenic capacity (Sana et al., 2018). Of the nine upregulated miRNA, miR-652, miR-345, and miR-9* all positively contributed to a higher risk score and poorer prognosis, while miR-301, miR153, miR-93, and miR-106b negatively contributed (Sana et al., 2018). These results signify that miRNAs are closely involved in the biological characteristics in GSCs, making them ideal targets for therapeutic treatment. MicroRNA Expression as a Biomarker The increase of certain microRNA strands allows them to be used as a biomarker by comparing their levels in the supposed cancerous tissue to levels in a known normal tissue of the same type. More research is needed until miRNA can act as a useful and reliable biomarker. The benefits of uncovering this diagnostic method are undoubtedly great as evident in breast cancer (BC), which already has research into its microRNA being reviewed as a possible diagnostic tool (Adhami et al., 2017). For miRNA to be used as a biomarker, there must be actual upregulation and downregulation of the different strands in cancerous tissue when compared to normal tissue. Studies show there are around 144 different miRNAs found with 74 being upregulated and 70 being downregulated (Adhami et al., 2017). In total, there are at least 30 differentially expressed miRNA across multiple studies. These were all expressed in a constant direction (Adhami et al., 2017) making them useful for use as a biomarker. Using this kind of detection shows great promise for modern day medicine. The current mammography technique is useful for catching early-stage breast cancer, but the technology is still unreliable and can lead to false positives as well as overdiagnosis and subsequent overtreatment of some minor cases (Adhami et al., 2017). And then there are those that do not get caught at all and are thus never treated. Using this new biomarker diagnosis via miRNA could help catch false positives or false negatives before any major harm is done. There are also a few miRNAs, specifically MiR-21, which have altered expression in many different types of human cancer (Adhami et al., 2017). Despite the potential of this study, there are still some drawbacks to using this kind of diagnostic marker. The first is the wide range of inconsistency found between research for most biomarker targets (Adhami et al., 2017). Second, researchers would need to compile a large library of all the consistently different regulations in order to be the most effective and accurate. Generally, the subject needs more research because of the major inconsistencies among studies and reviews regarding miRNA and its regulation (Adhami et al., 2017). MicroRNA as a Therapeutic Target MicroRNAs and their expression in glioblastoma cells may serve as a potential therapeutic target to treat this aggressive form of brain cancer. An overall lack of balance between tumor suppressor
Channels • 2021 • Volume 6 • Number 1 Page 5 miRNAs and oncomiRs contributes to the progression of various cancer types, and current researchers search for ways to correct the expression of certain microRNAs to possibly treat or slow the progression of cancer types. By understanding which miRNAs are differentially expressed in glioblastoma tumors and discovering how to control the expression of certain miRNAs, it may be possible to minimize or even reverse the effects of glioblastoma and lower the fatality rate of this type of brain tumor. When tumor-suppressing microRNAs lose function, malignant tumors can form. As previously discussed, microRNAs are responsible for regulating gene expression, and cancer research primarily focuses on the expression of tumor suppressor proteins and proto-oncogenes. Tumor suppressor microRNAs act by inhibiting the mRNAs that code for oncogenes. In many malignant tumors, tumor suppressor miRNAs have significantly lost their function through mutations, epigenetic silencing, or mistakes in mRNA processing (Mollaei et al., 2019). Some examples of a loss of function of tumor suppressor miRNAs in malignant tumors are miR-449a and miR495. MiR-449a is known to be downregulated in glioblastoma cells and may be contribute to the aggressive growth and multiplication of these cells. MiR495, on the other hand, is known to be downregulated in breast cancer, prostate cancer, and leukemia (Mollaei et al., 2019). Among many others, these downregulated miRNAs play a role in the development and progression of various cancer types. Another type of miRNA that plays a role in the development of cancer types is oncomiRs. OncomiRs are typically upregulated in cancerous tumor cells because of mutation or misregulation of certain pathways such as histone methylation or promoter methylation (Mollaei et al., 2019). Common miRNAs that are upregulated in cancer cells are miR-155 and miR-21. MiR-155 is overexpressed in lung cancer, breast cancer, and multiple forms of lymphoma, while MiR-21 is overexpressed in almost every type of human cancer—including blood and organ cancers (Mollaei et al., 2019). While little is known about the effects of upregulated miRNAs, it is clear they are important to the development of cancer like tumor suppressor miRNAs are. Understanding the expression of miRNAs and their role in cancer development could lead to development of a treatment for some of the most lethal cancers, such as glioblastoma. Theoretically, upregulating the downregulated tumor suppressor miRNAs or downregulating the upregulated miRNAs could recreate a balance between tumor suppressor proteins and oncogenes. Two current theories exist to explain how this information could be used in a viable treatment: miRNA reduction therapy and miRNA restoration therapy. MiRNA reduction, also called inhibition therapy, is based on the concept of inactivating upregulated miRNAs. Several methods are currently being used to try to meet this end, including miRNA sponges, locked-nucleic-acid antisense oligonucleotides, anti-miRNA oligonucleotides, miRNA nanoparticles, antagomirs, and multiple-target anti-miRNA antisense oligodeoxyribonucleotides (Mollaei et al., 2019). The basis behind all these methods is the same: to inhibit the expression of oncomiRs and slow the progression of various cancer types. However, there are some challenges related to this therapy. For example, many oncomiRs have multiple targets and focusing on limiting one miRNA at a time would not have a noticeable effect on cancer in a clinical setting (Mollaei et al., 2019). The concept behind the other method, miRNA restoration therapy, is to upregulate the tumor suppressor miRNAs and allow them to slow the production of oncoproteins within cancerous cells. The goal of this kind of therapy is to have cancerous cells take up microRNA genetic material that
Page 6 Hines, Marcum, Strong, Wade • Differential MicroRNA Expression will prevent the cells from continuing to grow and even cause an increase of apoptosis (Mollaei et al., 2019). After discussing the strategies behind using miRNAs to treat cancer types, it is important to understand how these theoretical therapies could be integrated into clinical medicine. Currently, biomedical researchers are investigating four main methods of drug delivery: anti-miRNA nucleotides, viral delivery, nonviral delivery, and small molecule drugs (Lee et al., 2020). The first type of delivery is the anti-miRNA nucleotides. RNA molecules are made more chemically stable by altering their sugar-phosphate background, which makes them less susceptible to degradation within the cell. By making RNA molecules more stable, certain nucleotide sequences could be synthesized and used to create anti-miRNA oligonucleotides that decrease the levels of specific miRNAs. Studies in mice have shown that this RNA modification can be successfully used to target a certain miRNA and decrease its expression within the cell in vivo (Lee et al., 2020). The second delivery strategy, viral delivery, has been studied for years as a mechanism to edit genes and change gene expression. In the case of cancer treatment, viruses can be used to control gene expression and limit harmful side effects, and viruses may also be able to remove certain destructive genes completely. For instance, a drug called Luxturna uses viral delivery to alter gene expression and treat Huntington’s disease by downregulating the Huntington gene (Lee et al., 2020). While this has not been used in clinical settings with cancer, it shows promising results as a therapeutic treatment. Nonviral delivery, the third method for delivery of treatment, includes polymeric vectors, lipids, and both inorganic and RNA nanoparticles. The goal of nonviral delivery is to allow the body to take up the drug without eliciting any immune response or causing the miRNAs to become completely degraded (Lee et al., 2020). The nonviral delivery methods described above have been proven to be safer for patients when compared to viral delivery, minimizing harmful side effects. The final delivery strategy that could be used to target miRNAs is the use of small molecule drugs. These drugs can be used to target specific miRNAs and transcription factors, changing the level of expression by inhibiting or inducing them (Lee et al., 2020). Small molecule drugs appear to be safe and effective for clinical use and may be able to help regulate the miRNA expression in a way that slows cancer progression in patients. Conclusion Summary of Findings Based on the way microRNAs (miRNAs) are upregulated or downregulated in cancerous tumors, specifically glioblastoma, there are two types of therapy that are being explored as potential treatments—miRNA reduction therapy and miRNA restoration therapy. MiRNA reduction therapy targets upregulated miRNAs and downregulates them in cancer patients to slow the progress of malignancies. MiRNA restoration therapy theoretically upregulates downregulated tumor suppressor miRNAs to also help slow cancer progression. Biomarkers can be used for diagnosing cancerous tumors. This is done by finding the specific miRNAs that are upregulated and downregulated in comparison with normal health brain tissue.
Channels • 2021 • Volume 6 • Number 1 Page 7 Using this sort of diagnostic can help reduce testing when a tumor can either be confirmed or ruled out. Potential Limitations As mentioned previously, there are a few limitations to the therapies that are currently being researched. First, it would be difficult to create a miRNA targeting treatment that the human body will accept; otherwise, the immune system would attack the treatment. Second, targeting one or two miRNAs would have minimal effect on cancerous tumors, and upregulating or downregulating a large number of miRNAs has proven to be challenging. Furthermore, when using them for detection, finding biomarkers in any patient is extremely difficult. Because multiple different markers exist, researchers would also need to compile a large library of marker types and compare those markers across healthy and cancerous tissue. Until these things are done, this detection method cannot be considered viable. Further Research Needed To use miRNAs as a potential treatment for glioblastoma, much additional research is needed. Aside from arranging clinical trials and developing a treatment that is safe and effective for clinical use, it is important to know exactly which miRNAs are impacted by glioblastoma and what their regulation patterns are. Researchers would also need to gain a better understanding for what microRNAs cause glioblastoma and which microRNAs are affected as a result of glioblastoma. Additional research on these topics is crucial to creating the most effective treatment. Many more tests of potential biomarkers also need to be done for the effective use of biomarkers. As mentioned, a large library of biomarkers must be built up. Then, researchers need to find consistency in each type for them to be used as biomarkers for possibly presenting a cancerous tumor. Once consistency is established, scientists must determine how to administer the tests in the least invasive, most reliable, and most efficient way possible. Then, the full potential of the biomarkers can be used to help detect the cancer while it is still treatable. Developing a broad understanding of microRNAs and the role they play in autoimmune disorders and cancer, specifically neural cancers, can lead to advancements in the field of molecular biology research. MicroRNAs were recently discovered, and there is still much to learn about their expression, purpose, and limitations as it relates to various diseases. As more experimental data emerges, the hypotheses that scientists now have regarding microRNAs may be accepted or changed, shaping future research, treatments, and diagnostic procedures.
Page 8 Hines, Marcum, Strong, Wade • Differential MicroRNA Expression Bibliography Adhami, M., Haghdoost, A. A., Sadeghi, B., & Malekpour Afshar, R. (2017). Candidate miRNAs in human breast cancer biomarkers: a systematic review. Breast Cancer, 25(2), 198–205. https://doi.org/10.1007/s12282-017-0814-8 Colamatteo, A., Micillo, T., Bruzzaniti, S., Fusco, C., Garavelli, S., De Rosa, V., . . . Matarese, G. (2019). Metabolism and autoimmune responses: The microRNA connection. Frontiers in Immunology, 10. Retrieved February 24, 2021, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722206/ Diana, Andrea, et al. “MicroRNAs at the Crossroad of the Dichotomic Pathway Cell Death vs. Stemness in Neural Somatic and Cancer Stem Cells: Implications and Therapeutic Strategies.” International Journal of Molecular Sciences, vol. 21, no. 24, 2020, p. 9630., doi:10.3390/ijms21249630. Gimple, R. C., Bhargava, S., Dixit, D., & Rich, J. N. (2019). Glioblastoma stem cells: lessons from the tumor hierarchy in a lethal cancer. Genes & development, 33(11-12), 591–609. Retrieved February 16, 2021, from https://doi.org/10.1101/gad.324301.119 Lafourcade, Carlos, et al. “MIRNAS in Astrocyte-Derived Exosomes as Possible Mediators of Neuronal Plasticity.” Journal of Experimental Neuroscience, vol. 10s1, 2016, doi:10.4137/jen.s39916. Lee, T. J., Kerr, K., Barker, E. L., Eltzschig, H. K., Kaur, B., Kim, D. H., . . . Yuan, X. (2020). Strategies to Modulate MicroRNA Functions for the Treatment of Cancer or Organ Injury. ASPET Pharmacological Reviews, 72(3). Retrieved February 26, 2021, from https://pharmrev.aspetjournals.org/content/72/3/639.long#sec-7 Liu, C., Yang, H., Shi, W., Wang, T., & Ruan, Q. (2018). MicroRNA-mediated regulation of T helper type 17/regulatory T-cell balance in autoimmune disease. Immunology, 155(4). Retrieved February 24, 2021, from https://pubmed.ncbi.nlm.nih.gov/30133700/ Mollaei, H., Safaralizadeh, R., & Rostami, Z. (2019). MicroRNA replacement therapy in cancer. Journal of Cellular Physiology, 234(8). Retrieved February 26, 2021, from https://onlinelibrary.wiley.com/doi/10.1002/jcp.28058 Sana, J., Busek, P., Fadrus, P., Besse, A., Radova, L., Vecera, M., Reguli, S., Stollinova Sromova, L., Hilser, M., Lipina, R., Lakomy, R., Kren, L., Smrcka, M., Sedo, A., & Slaby, O. (2018). Identification of microRNAs differentially expressed in glioblastoma stem-like cells and their association with patient survival. Scientific reports, 8(1), 2836. Retrieved February 18, 2021, from https://doi.org/10.1038/s41598-018-20929-6 Vijayaraghava T.S. Rao, PhD, Samuel K. Ludwin, MD, Shih-Chieh Fuh, BSc, Robin Sawaya, BSc, Craig S. Moore, PhD, Ming-Kai Ho, MSc, Barry J. Bedell, MD, PhD, Harvey B. Sarnat, MD, Amit BarOr, MD, Jack P. Antel, MD, MicroRNA Expression Patterns in Human Astrocytes in Relation to Anatomical Location and Age, Journal of Neuropathology & Experimental Neurology, Volume 75, Issue 2, February 2016, Pages 156–166, https://doi.org/10.1093/jnen/nlv016
Channels • 2021 • Volume 6 • Number 1 Page 9 Channels Vol. 6 No. 1 (2021): 9–20 ISSN 2474-2651 © 2021, Emily Walker, licensed under CC BY-NC-ND (http://creativecommons.org/licenses/by-nc-nd/4.0/) Humor in the Foreign Language Classroom Emily Walker English, Literature, and Modern Languages Introduction oreign language (FL) pedagogy has seen much evolution as language instruction research has progressed from the grammar-translation methods to the audio-lingual, then to the current emphasis on communicative competence. As researchers study the components that make an effective FL classroom, they strive to formulate a concept of “best practice.” Toward this end, this research examines the effects of humor on one college-level FL class through qualitative data collection and synthesis of the themes that present themselves during the study. Literature Review Preliminary Key Constructs One factor that must be addressed is the affective filter. As presented by Krashen (1981) and explained by Du (2009), the affective filter hypothesis posits a mental block that prevents comprehensible input from being used for language acquisition. Factors influencing the affective filter include student motivation, attitude, anxiety, and self-confidence (Du, 2009). For the purposes of this research, I’ve also restricted the discussion of pedagogical tools to resources that support and enhance teaching. In referring to foreign language (FL) classrooms, I wish to demarcate mainly classrooms in which students learn a language that is not commonly used in the country where they live. Secondary Key Constructs In accordance with the findings later presented, it is necessary to address the constructs of power distance and willingness to communicate (WTC). As introduced by Hofstede (2011) in his seminal categorization of selected constructs in intercultural communication, power distance is “the extent to which the less powerful members of organizations and institutions (like the family) accept and expect that power is distributed unequally” (p. 9). Specifically, power distance is the perceived legitimacy of hierarchy within a social organization as defined not only by those at the upper levels, but also by those in the lower ranks of the pyramid. This study focuses on the level of informality in the classroom and the approachability of the professor as key elements of power distance. WTC, on the other hand, is characterized by the intersection of students’ perceived communicative competence and their level of anxiety in the FL classroom (Yashima, 2002). Students with lower anxiety who believe themselves to have a greater ability to communicate in the FL classroom will
Page 10 Walker • Humor in the Foreign Language Classroom typically be more willing to speak up in class and practice using the L2. The constructs of WTC and power distance will be used to frame much of the Findings and Discussion section. Approach to Humor In order to examine the discussion of humor in the FL classroom and its effect on students’ affective filters, we must first broach the construct of humor itself. Because humor is difficult to define, many previous studies have relied on readers’ intuitive understanding of what humor is (Bell, 2009). A few trends can be identified in what we recognize as humor: subverting expectations, such as violating Gricean maxims; politeness conventions; social taboos; joking (not canned jokes, but interactional humor and joking with students); exaggeration and sarcasm; and self-deprecation (Azizifard & Jalali, 2012; Dynel, 2009). Nevertheless, Bell (2009) warns that while these typologies might be a good starting point for understanding varieties of humor, humor is complex and these categorizations “cannot be taken as accurately representing conversational joking” (p. 244). If meticulous typologies of humor are thus deemed inadequate and intuition remains a major categorizing factor, we must look at how previous researchers have gone about studying it in classrooms. Beginning with a common framework in educational research, Askildson (2005) categorizes his discussion according to direct and indirect effects of humor—the former affecting the saliency of input and information retention and the latter influencing the general class environment and factors of the affective filter. While the direct effects have been well analyzed through experimental testing and detailed qualitative research, the indirect effects have only recently begun to receive attention in literature, perhaps due to their less defined nature (Zabidin, 2015). In his 2010 article on interactional humor, Norrick discusses indirect effects of humor, noting that humor “contributes to the creation of identity and fosters group rapport” (p. 240). He also notes that joking works as “positive politeness” that builds camaraderie by lessening the distance between speakers and reducing the threat of impositions on the listener. Furthermore, he addresses an important construct—how the framing of actions in a sociolinguistic context determines the reactions they will elicit. Linguistic and paralinguistic features can signal a “play” context rather than a serious one, setting the stage for the entire interaction. Within a play frame, even insulting, mocking, or otherwise impolite speech acts can be received good-naturedly. Facework also becomes relevant here as joking can be used to reduce the threat of impositions on the listener and save face for either interlocutor (Norrick, 2010). Further in his research, Askildson (2005) describes humor as “a pedagogical instrument like any other, and one which serves as a double-edged sword—capable of improving or harming the classroom environment depending on its employment by the teacher” (p. 48-49). He notes the possible negative effects of offense and confusion if humor is used improperly or in a poor context. In his own study, Askildson aimed for a representative sample of perspectives by using a Likertscaled questionnaire to target students in various language classes. Most of the participants reported that they felt either noticeably or considerably more relaxed in class when humor was employed, and students perceived teachers as more approachable when they used humor. Participants also seemed to view target language humor as an important tool in language and cultural learning.
Channels • 2021 • Volume 6 • Number 1 Page 11 Classroom language in general presents an interesting context for humor research. In their 2014 publication, Bell and Pomerantz argue that much of formal language instruction is a fiction, or more accurately a fabrication, as teachers and learners are aware that most classroom language varies significantly from authentic communication among native speakers. Humor seeps into this crack, as stylistic variation and non-serious talk are ubiquitous in real-world interactions. Although Bell and Pomerantz are not the first to criticize the constraints of language education, their paper begins to fill the gap in applying alternative conceptions of language to L2 pedagogy, increasing the focus on interaction and communicative competency. Notably, they indicate that “humor and language play allow for and even require learners to adopt a view of communication that is predicated on joint negotiation, emergence of meaning, and mediation” ((Bell & Pomerantz, 2014, p. 40). Using humor to emphasize these factors in a language classroom encourages learners to creatively use both communicative and interpretive resources as they interact in the classroom. Honing in on the use of L1 humor in FL classrooms, Askildson (2005) observes, “The overwhelming majority of those surveyed indicated that even general (non-target language) humor was an important element of creating an overall environment conducive to learning. Specifically, participants indicated reduced anxiety/tension, improved approachability of teachers, and increased levels of interest as a result of humor usage by the teacher” (p. 55). Notably, reduced anxiety and motivation are associated with the lowering of the affective filter. Askildson goes on to state that humor’s perceived importance in learning should guide our consideration of the concept in research on pedagogy (p. 56). Finally, Askildson notes that “humor’s evident ability to lower the affective filter makes a strong argument in and of itself for explicit inclusion of humor in a language educational context” (p. 49). Given the results of his and others’ research in the field, Askildson strongly advocates further research on the pedagogical effects of humor in language-specific classrooms. Researchers Ziyaeemehr and Kumar (2014) also address a few aspects of humor’s role in the FL classroom, particularly its role in “foregrounding form,” “highlighting cultural dissimilarities,” and the notion of a “play frame,” similar to Norrick’s (2010) construct. They emphasize highlighting content and drawing students’ attention through the use of humor. That usage brings cultural and linguistic dissimilarities into relief thanks to contrasting expectations. Additionally, putting recasts of errors and more difficult language distinctions into a play frame politely corrects students’ errors and draws their attention to the particular structure in question (Ziyaeemehr & Kumar, 2014, p. 7). These features form a framework for analyzing the direct and indirect effects of humor in an FL classroom. Lastly, Bilokcuoglu and Debreli (2018) tout the potential of humor in “creating an affirmative environment and for establishing a less authoritarian way of teaching, leading to the reduction of affective filter among learners” (p. 356). Their article points out that given the prevalence of humor in everyday interactions and the potential of humor in classroom settings, it is surprising that more research has not been done on how to best use humor as a motivational and tension-reducing tool. Humor allows teachers to assume the role of facilitators instead of high-authority lecturers, especially in communicative classrooms. Often, teachers view humor as a distraction or cause of chaos rather than a tool for effective learning. Used correctly, humor can foster a sense of belonging for students and allow them to participate without losing face. These ideas must be balanced with the potential negative effects of overusing humor. In particular, sarcastic humor can lead to confusion in the L2, and humor directed too frequently at a single person might be misinterpreted
Page 12 Walker • Humor in the Foreign Language Classroom and produce negative effects, depending on the nature of the humor. Bilokcuoglu and Debreli (2018) conclude by stressing the need for more research on humor in language classroom settings. A Particular Need Because this is still a growing niche in the field of linguistics, much research remains to be done about specific classroom situations in which humor is used (Askildson, 2005; Bell & Pomerantz, 2014; Bilokcuoglu & Debreli, 2018). As it stands, a disconnect remains between theory and studies. Much of the existing research consists of either larger questionnaire-based studies like Askildson’s, which provide more quantitative findings, or simply theoretical discussions on humor’s potential in language classrooms. As these researchers make evident, case study research in actual classroom situations is necessary to invigorate the academic conversation concerning the effects of humor usage. Within this context, I present my research question as follows: How does L1 humor, specifically when used as a pedagogical tool, influence students’ affective filters in a college-level elementary foreign language classroom? Methods This research, framed as a case study of a college-level elementary French classroom, was comprised of classroom observations and student interviews for the purpose of triangulation (Harbon & Shen, 2010). The beginning French class, offered at a small liberal arts college in the midwestern United States, consisted of eight college-aged students and one professor. Due to the school’s COVID-19 protocol in the fall 2020 semester, full class attendance was occasionally disrupted by student quarantines or absences. Despite this, students were usually able to participate in class over Zoom, and I was able to collect sufficient data from the remaining in-person students. In order to formulate a rough observation scheme (Harbon & Shen, 2010), I began by conducting a preliminary observation of the French class to provide a framework for the later observations. Harbon and Shen (2010) suggest that researchers may wish to seek a balance between an observation scheme and field notes in order to include aspects that a structured protocol might not capture. Consequently, I collected both field notes (loosely guided by my preliminary observation) and video recordings of four fifty-minute class sessions throughout several weeks in the middle of the fall semester. To uphold an unobtrusive, non-judgmental yet emic observation style, I recorded these class meetings from a back corner of the classroom. I did not participate in class activities but introduced myself before the first observation and made friendly conversation with the students before and after class to make them comfortable with my presence in the classroom. This posture was indubitably aided by my own membership as a student at the school. My repeated presence in the class also reduced observer effect (Labov, 1972). Of the eight students in the class, four consented to be interviewed, and each interview lasted approximately forty minutes. The development of interview questions was guided by an emphasis on co-construction and reflexivity (Mann, 2011), as well as reflection on the collected data. The interview protocol for this study (see Appendix A) consisted of questions designed to assemble an interviewee profile and elicit their perspectives on classroom humor (Wagner, 2010). I began each interview by explaining the concept and goals of the study and the student’s role as an interviewee,
Channels • 2021 • Volume 6 • Number 1 Page 13 then initiating simple conversation to set them at ease. Next, I transitioned to questions concerning their language experience, humor preferences, and general attitudes. We continued with a discussion of the participant’s perspectives on class humor in general as well as specific instances from the class periods (Harbon & Shen, 2010; Wagner, 2010). Throughout the interviews, I intentionally paraphrased the interviewee’s words back to them and directly invited validation or correction of the stated concepts to ensure accuracy as a form of member checking. Due to this emphasis on co-construction and natural elicitation of data (Mann, 2011), each interview naturally followed a slightly different train of conversation. The questions listed in the interview protocol were mere starting points, as the goal of the interviews was to begin a conversation around the context to gain a more holistic understanding of the situation (Harbon & Shen, 2010). Finally, as suggested by Mann (2011), I embraced my own interactional influence and potential bias as a peer researcher, student, and fellow language learner, using these as an opportunity to elicit student responses in a manner that researchers of other backgrounds might not be able to. I obtained approval from the university’s Institutional Review Board to conduct this study. The process of data analysis was greatly shaped by acknowledging my underlying assumptions as a student researcher and adding another level of reflexivity to mitigate unwanted skew (Mann, 2011; Wagner, 2010). My method of data collection was informed by Holiday (2010); based on his suggestions, I began with broad observations, then moved to personalized thick descriptions, and landed on focused inquiry. My approach was influenced by transparency in method, submission to data, and dedication to making appropriate claims in order to maintain reliability (Holliday, 2010). According to the suggestions of Wagner (2010), I also considered the effects of self-deception bias and acquiescence bias on participants’ responses to interview questions. In analysis, I strove to let the research be driven by the themes the data elucidated. To draw proper conclusions from subjective data, I coded my transcribed interviews and portions of the video recordings from observations, then grouped them according to themes. Using these themes, I formed a tentative argument that I revised as I continually refined my data analyses and gradually approached a formal set of findings (Holliday, 2010). Finding and Discussions Reflection on the Research Question How does L1 humor, specifically when used as a pedagogical tool, influence students’ affective filters in a college-level elementary foreign language classroom? During the course of this study, diverse themes emerged from the data beyond what I had originally anticipated in my research question. Originally, I expected to gather information concerning humor as a pedagogical tool and its impact on the affective filter. However, no students reported seeing humor as a tool; on the contrary, Participant 1 noted, “I do think a lot of this is just her personality. I do think she likes to make us laugh, but I don’t think she goes out of her way to make us laugh.” The other three participants expressed similar impressions, commenting that the professor’s style of humor seems to be a natural extension of her personality. I can report little on the affective filter element, which I had used as a framing device for my research question. The data gathered does not
Page 14 Walker • Humor in the Foreign Language Classroom emphasize the affective filter element as expected. Rather, as I questioned my participants, three themes emerged: power distance, self-effacing humor, and willingness to communicate (WTC). These constructs, as a natural outpouring of my data, have consequently become the focus of my findings. Power Distance The concept of power distance, though less cited in language instruction research than in intercultural communication studies, is certainly at play in this French classroom (Hofstede, 2011). Power distance appears to be largely tied to class environment in this situation; the professor’s low power distance structure in class was evident in the field observations, and all four participants reported class factors related to lower power distance. Participant 1 introduced the concept of power distance unprompted, noting, “She’s…to refer to it in intercultural terms, very low power distance structure. Which works, because I have a very low power distance structure.” He further described the professor’s teaching style as “easygoing, very informal.” Participant 2 affirmed that her lighthearted humor makes the classroom very relaxed, saying, “It’s a main contributor to her classroom environment, that it can be so open—she makes jokes about herself.” Participants 3 and 4 brought up the professor’s graceful response to confrontational questions, emphasizing that she does not see correction as a threat to her authority. Participant 4 also discussed how the professor trusts and respects her students enough to give them freedom in the classroom and with homework. These findings align with Tananuraksakul’s (2013) observations that “breaking down the high degree of PD [power distance] can help boost the affective sides to some degrees” (p. 112). In this manner, the findings of lowered power distance in the classroom are still tied to the construct of the affective filter. Many of the same factors are intertwined here, including a relaxed atmosphere and more individual freedom. Self-Effacing Humor In many ways, the power distance in this classroom was affected by the professor’s particular brand of humor. Labeled “self-effacing” by Students 1 and 4, her humor is characterized by lighthearted exaggeration, usually at her own expense. According to Participants 1 and 2, it often feels spontaneous or situational—“opportunistic,” according to Student 1. The professor makes constructive use of “teachable moments” built on her own slips in class. Participant 4 mentioned that this conveys the mentality that mistakes are ways to learn. Through classroom observations, it was also evident that the professor subtly shapes their perspective of communication in the FL context by lessening the gravity surrounding language errors and presenting them as continuous opportunities instead of fatal failures. One example of this professor’s humor, noted during the observational stage of the research, involves a poorly drawn stick-figure. As part of a vocabulary illustration, the professor attempted to draw a person on the whiteboard, but many of the limbs were obviously disjointed and the head was floating ominously above the body. As the students started to chuckle at the figure, which appeared to need medical attention, the professor quickly looked back at the class and declared, “Je suis artiste! [I am an artist!]” with a dramatic wave of her hand.
Channels • 2021 • Volume 6 • Number 1 Page 15 This sort of arrogance, exaggerated with a grin, is well-tempered by the professor’s quick willingness to admit her own mistakes and share embarrassing stories about her language gaffes. “She sometimes talks about her own faux paus and…mistakes she had in French, stories with her husband’s family,” noted Participant 3. “She’s comfortable making fun of herself.” This sharing also contributes to the perception of low power distance in the classroom, instituting a sense of relatability and shared experience with the students. These frequent airy asides contribute to what Norrick (2010) would label a “play” context, allowing students to experience every part of the class in a less threatening manner. When prompted to discuss how they perceived their professor’s humor, three of the four participants made sure to clarify that humor was not a distraction within the classroom, but rather a beneficial and engaging aspect that increased their level of comfort in class. Willingness to Communicate Each of the aforementioned factors affects students’ WTC, an overarching theme highlighted by the interviewed students. The lowered power distance and use of humor both contribute to an affirmative class environment. All four participants stated that the professor’s humor contributed to an open environment in which they felt more comfortable practicing the language. As Participant 3 put it, “She puts people at ease with her humor and makes situations easier to be in.” By laughing at her own mistakes, the professor shows students that they can make mistakes too. She also tries to break things down when they seem difficult. “She’ll just make some lighthearted comments as she’s writing things down on the board…she makes the language feel more manageable,” Participant 2 explained. All four participants noted that they felt very comfortable and free to try speaking in the class, despite the risk of making mistakes. Even those who admitted they were not yet incredibly confident in their language skills reported this feeling of comfort in the French classroom. As Student 3 attested, “I’m not a person who would raise my hand or speak out in class, but it’s easier to do that in her class.” Likewise, Participant 2 noted that humor keeps her engaged in the class, increasing her WTC because of the interaction with and sense of personal investment from the professor. Additional Themes Another tangential theme elucidated by the interviews was the professor’s care for her students. Participant 4 established that making the effort to use humor shows that the professor is more engaged in the class herself, using it to check in on students. “If the teacher seems uninterested or stiff…how are the students supposed to pay attention or be interested?” she asks. Participant 3 affirms this, saying, “When the professor cares, it’s easier for you to care.” Students also cited this personal attention as a contributing factor to WTC. Limitations & Further Research Some of the major limitations of this study were the short time span allotted for data collection, the limited number of participants, students’ perceptions of me as a researcher, and of course my own biases. The restrictions of this study allowed for only a few observation sessions collected over a limited period of time and are not necessarily representative of the class dynamic throughout the
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